Temporal and spatial transcriptional programs in murine kidney development
Temporal and spatial transcriptional programs in murine kidney development
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DOI：
10.1152/physiolgenomics.00043.2005
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发表时间：
2005-10-17
影响因子：
4.6
通讯作者：
Grimmond, SM
中科院分区：
文献类型：
--
作者：
Challen, G;Gardiner, B;Grimmond, SM
我们已经进行了系统的时间和空间的表达谱的发育小鼠肾脏使用Compugen长寡核苷酸微阵列。从出生后10.5天（dpc）后肾间充质（MM）到新生儿肾脏，以24小时的间隔监测18，000个基因的活性，并确定了3，600个动态表达基因的队列。通过直接比较来自10.5 vs. 11.5 vs. 13.5dpc肾的RNA进一步调查早期后肾发育。这些数据显示与先前公布的大鼠肾脏发育的动态概况（StuartRO，Bush KT，and Nigam SK. Proc Natl Acad Sci USA 98：5649-5654，2001）和我们自己的时间数据高度一致。聚类分析被用来确定基因本体论术语，功能注释，和与时间表达谱相关的途径。遗传网络分析也被用来确定在早期后肾发育过程中具有最大转录活性的生物网络，突出了增殖和分化的参与。利用分期胚胎肾脏的整体和切片原位杂交验证差异基因表达。还进行了两次空间特征分析实验。将MM（10.5dpc）与相邻中间间充质进行比较，以进一步确定后肾定位。为了定义参与分支和诱导肾发生的基因，在15.5dpc时从HoxB 7-GFP转基因小鼠分选的输尿管芽（GFP+）FACS上相对于GFP-间充质衍生物进行表达谱分析。时间和空间数据之间的比较增强了预测基因和网络功能的能力。这项研究提供了迄今为止最全面的肾脏发育的时间和空间调查，这些转录调查的汇编提供了重要的见解后肾发育，现在可以进行功能测试。
We have performed a systematic temporal and spatial expression profiling of the developing mouse kidney using Compugen long-oligonucleotide microarrays. The activity of 18,000 genes was monitored at 24-h intervals from 10.5-day-postcoitum (dpc) metanephric mesenchyme (MM) through to neonatal kidney, and a cohort of 3,600 dynamically expressed genes was identified. Early metanephric development was further surveyed by directly comparing RNA from 10.5 vs. 11.5 vs. 13.5dpc kidneys. These data showed high concordance with the previously published dynamic profile of rat kidney development (Stuart RO, Bush KT, and Nigam SK. Proc Natl Acad Sci USA 98: 5649-5654, 2001) and our own temporal data. Cluster analyses were used to identify gene ontological terms, functional annotations, and pathways associated with temporal expression profiles. Genetic network analysis was also used to identify biological networks that have maximal transcriptional activity during early metanephric development, highlighting the involvement of proliferation and differentiation. Differential gene expression was validated using whole mount and section in situ hybridization of staged embryonic kidneys. Two spatial profiling experiments were also undertaken. MM (10.5dpc) was compared with adjacent intermediate mesenchyme to further define metanephric commitment. To define the genes involved in branching and in the induction of nephrogenesis, expression profiling was performed on ureteric bud (GFP+) FACS sorted from HoxB7-GFP transgenic mice at 15.5dpc vs. the GFP- mesenchymal derivatives. Comparisons between temporal and spatial data enhanced the ability to predict function for genes and networks. This study provides the most comprehensive temporal and spatial survey of kidney development to date, and the compilation of these transcriptional surveys provides important insights into metanephric development that can now be functionally tested.